Arabidopsis abcbl abcb19 double mutants defective in the auxin transporters ABCB1/PGP1 and ABCB19/PGP19 are altered in stamen elongation, anther dehiscence and pollen maturation. To assess the contribution of these tr...Arabidopsis abcbl abcb19 double mutants defective in the auxin transporters ABCB1/PGP1 and ABCB19/PGP19 are altered in stamen elongation, anther dehiscence and pollen maturation. To assess the contribution of these transporters to stamen development we performed phenotypic, histological analyses, and in situ hybridizations on abcbl and abcbl9 single mutant flowers. We found that pollen maturation and anther dehiscence are precocious in the abcbl but not in the abcb19 mutant. Accordingly, endothecium ligniflcation is altered only in abcbl anthers. Both abcbl and abcb1 abcb19 stamens also show altered early development, with asynchronous anther Iocules and a multilayer tapetum. DAPI staining showed that the timing of meiosis is asynchronous in abcbl abcb19 anther Iocules, while only a small percentage of pollen grains are non- viable according to Alexander's staining. In agreement, TAM (TARDY ASYNCHRONOUS MEIOSIS), as well as BAM2 (BARELY ANY MERISTEM)involved in tapetal cell development--areoverexpressed in abcbl abcb19 young flower buds. Corre- spondingly, ABCB1 and ABCB19 mRNA localization supports the observed phenotypes of abcbl and abcbl abcb19 mutant anthers. In conclusion, we provide evidence that auxin transport plays a significant role both in early and late stamen development: ABCB1 plays a major role during anther development, while ABCB19 has a synergistic role.展开更多
Cullin-RING E3 ligases (CRLs) regulate different aspects of plant development and are activated by modification of their cullin subunit with the ubiquitin-like protein NEDD8 (NEural precursor cell expressed Develop...Cullin-RING E3 ligases (CRLs) regulate different aspects of plant development and are activated by modification of their cullin subunit with the ubiquitin-like protein NEDD8 (NEural precursor cell expressed Developmentally Down-regulated 8) (neddylation) and deactivated by NEDD8 removal (deneddylation). The CONSTITUTIVELY PHOTOMORPHOGENIC9 (COP9) signalosome (CSN) acts as a molecular switch of CRLs activity by reverting their neddylation status, but its contribution to embryonic and early seedling development remains poorly characterized. Here, we analyzed the phenotypic defects of csn mutants and monitored the cullin deneddylation/neddylation ratio during embryonic and early seedling development. We show that while csn mutants can complete embryogenesis (albeit at a slower pace than wildtype) and are able to germinate (albeit at a reduced rate), they progressively lose meristem activity upon germination until they become unable to sustain growth. We also show that the majority of cullin proteins are progressively neddylated during the late stages of seed maturation and become deneddylated upon seed germination. This developmentally regulated shift in the cullin neddylation status is absent in csn mutants. We conclude that the CSN and its cullin deneddylation activity are required to sustain postembryonic meristem function in Arabidopsis.展开更多
基金partially supported by a research grant from MIUR (PRIN) to P.C. and M.C.a research grant to M.C. from the Italian Ministry of Foreign Affairs (Direzione Generale per la Promozione del Sistema Paese, Unità per la cooperazione scientifica e tecnologica bilaterale e multilaterale)
文摘Arabidopsis abcbl abcb19 double mutants defective in the auxin transporters ABCB1/PGP1 and ABCB19/PGP19 are altered in stamen elongation, anther dehiscence and pollen maturation. To assess the contribution of these transporters to stamen development we performed phenotypic, histological analyses, and in situ hybridizations on abcbl and abcbl9 single mutant flowers. We found that pollen maturation and anther dehiscence are precocious in the abcbl but not in the abcb19 mutant. Accordingly, endothecium ligniflcation is altered only in abcbl anthers. Both abcbl and abcb1 abcb19 stamens also show altered early development, with asynchronous anther Iocules and a multilayer tapetum. DAPI staining showed that the timing of meiosis is asynchronous in abcbl abcb19 anther Iocules, while only a small percentage of pollen grains are non- viable according to Alexander's staining. In agreement, TAM (TARDY ASYNCHRONOUS MEIOSIS), as well as BAM2 (BARELY ANY MERISTEM)involved in tapetal cell development--areoverexpressed in abcbl abcb19 young flower buds. Corre- spondingly, ABCB1 and ABCB19 mRNA localization supports the observed phenotypes of abcbl and abcbl abcb19 mutant anthers. In conclusion, we provide evidence that auxin transport plays a significant role both in early and late stamen development: ABCB1 plays a major role during anther development, while ABCB19 has a synergistic role.
文摘Cullin-RING E3 ligases (CRLs) regulate different aspects of plant development and are activated by modification of their cullin subunit with the ubiquitin-like protein NEDD8 (NEural precursor cell expressed Developmentally Down-regulated 8) (neddylation) and deactivated by NEDD8 removal (deneddylation). The CONSTITUTIVELY PHOTOMORPHOGENIC9 (COP9) signalosome (CSN) acts as a molecular switch of CRLs activity by reverting their neddylation status, but its contribution to embryonic and early seedling development remains poorly characterized. Here, we analyzed the phenotypic defects of csn mutants and monitored the cullin deneddylation/neddylation ratio during embryonic and early seedling development. We show that while csn mutants can complete embryogenesis (albeit at a slower pace than wildtype) and are able to germinate (albeit at a reduced rate), they progressively lose meristem activity upon germination until they become unable to sustain growth. We also show that the majority of cullin proteins are progressively neddylated during the late stages of seed maturation and become deneddylated upon seed germination. This developmentally regulated shift in the cullin neddylation status is absent in csn mutants. We conclude that the CSN and its cullin deneddylation activity are required to sustain postembryonic meristem function in Arabidopsis.